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bioRxiv · 10.1101/2020.11.24.396507

Protein Detection and Localization in Plant Cells Using Spot-Tagging

Mentzelopoulou, A.·Liu, C.·Moschou, P.
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Abstract

Fluorescent labelling of proteins without compromising their activity is crucial for determining their spatiotemporal localization while retaining their functionality. Spot-tag is a 12-amino acid peptide recognized by a single-domain nanobody. Here we introduce the spot-tag as a labelling strategy for proteins in fixed and living plant cells, using as an example the microtubule motor centromeric protein E-related Kinesin 7.3. Spot-tagging of ectopically introduced Kinesin 7.3 does not interfere with microtubules and spot staining results in a close-grained fluorophore labelling revealing a localization pattern that resembles "beads-on-a-string". We anticipate that our protocol will apply to many more demanding protein cellular targets, offsetting activity perturbations and low photon quantum yields imposed by other protein-tagging approaches.

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Mentzelopoulou, A., Liu, C., Moschou, P.. 2020-11-24. Protein Detection and Localization in Plant Cells Using Spot-Tagging. https://doi.org/10.1101/2020.11.24.396507

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